Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Neuron transfection method-need the best protocol


  • Please log in to reply
4 replies to this topic

#1 katenkak

katenkak

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 52 posts
0
Neutral

Posted 23 March 2011 - 01:43 AM

Topic says its all. I need to transfect mouse cerebellar and/or hippocampal neurons with plasmid or siRNA. What works best with highest survival rate? I have tried Amaxa electroporation-neurons look ugly and don't survive. When I tried AMaxa with plasmid, neurons survived for 3 days and died. I try with Lipofectamine 2000 and see that during 4 hours of incubation with Lipofectamine, many neurons die and processes of remaining neurons get disintegrated. Moreover efficiency of transfection is close to 1% contra promised 25-20% . Please help me if you have a protocol!!!! Thank you!

#2 HBImolbiol

HBImolbiol

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 23 March 2011 - 07:32 AM

Topic says its all. I need to transfect mouse cerebellar and/or hippocampal neurons with plasmid or siRNA. What works best with highest survival rate? I have tried Amaxa electroporation-neurons look ugly and don't survive. When I tried AMaxa with plasmid, neurons survived for 3 days and died. I try with Lipofectamine 2000 and see that during 4 hours of incubation with Lipofectamine, many neurons die and processes of remaining neurons get disintegrated. Moreover efficiency of transfection is close to 1% contra promised 25-20% . Please help me if you have a protocol!!!! Thank you!


You could try a viral vector such as Adeno associated virus (serotype 9) or lentivirus.

#3 katenkak

katenkak

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 52 posts
0
Neutral

Posted 23 March 2011 - 07:34 AM

1) Do you know a best vector for lentiviral expression in neurons?
2) I try to find a non-viral protocol, since according to local rules, I can't take live cells outsinde og class 2 area, but only fixed. I need live neurons for real-time measurements.

Edited by katenkak, 23 March 2011 - 07:36 AM.


#4 swimmer

swimmer

    member

  • Members
  • Pip
  • 1 posts
-1
Neutral

Posted 20 June 2011 - 06:51 AM

You may try to find sth. from this paper.
http://www.nature.co...bs/ncb1881.html

Edited by swimmer, 20 June 2011 - 06:52 AM.


#5 labrat612

labrat612

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 87 posts
1
Neutral

Posted 05 July 2011 - 06:31 AM

Have you tried Calcium-phosphate method? It's tedious and time-consuming but it does give descent results.


~Labrat612




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.